Intact-Cell MALDI-ToF Mass Spectrometry for the Authentication of Drug-Adapted Cancer Cell Lines
Jane F Povey1, Emily Saintas2, Adewale V Aderemi3
1Industry Biotechnology Centre and School of Biosciences, University of Kent, Canterbury CT2 7NJ, UK. J.Povey@kent.ac.uk.
Abstract:
The use of cell lines in research can be affected by cell line misidentification. Short tandem repeat (STR) analysis is an effective method, and the gold standard, for the identification of the genetic origin of a cell line, but methods that allow the discrimination between cell lines of the same genetic origin are lacking. Here, we use intact cell MALDI-ToF mass spectrometry analysis, routinely used for the identification of bacteria in clinical diagnostic procedures, for the authentication of a set of cell lines consisting of three parental neuroblastoma cell lines (IMR-5, IMR-32 and UKF-NB-3) and eleven drug-adapted sublines. Principal component analysis (PCA) of intact-cell MALDI-ToF mass spectrometry data revealed clear differences between most, but not all, of the investigated cell lines. Mass spectrometry whole-cell fingerprints enabled the separation of IMR-32 and its clonal subline IMR-5. Sublines that had been adapted to closely related drugs, for example, the cisplatin- and oxaliplatin-resistant UKF-NB-3 sublines and the vincristine- and vinblastine-adapted IMR-5 sublines, also displayed clearly distinctive patterns. In conclusion, intact whole-cell MALDI-ToF mass spectrometry has the potential to be further developed into an authentication method for mammalian cells of a common genetic origin.
Insights
Intact cell MALDI-ToF mass spectrometry can distinguish between cell lines, even those from the same genetic origin. This method shows potential for authenticating mammalian cell lines, addressing a key research challenge.
Area of Science:
- Biotechnology
- Cell Biology
- Analytical Chemistry
Background:
- Cell line misidentification poses a significant challenge in scientific research.
- Short tandem repeat (STR) analysis is the standard for cell line genetic origin identification but cannot differentiate sublines.
- A need exists for methods to authenticate cell lines with shared genetic origins.
Purpose of the Study:
- To evaluate intact cell MALDI-ToF mass spectrometry for differentiating neuroblastoma cell lines and their drug-adapted sublines.
- To assess the potential of MALDI-ToF mass spectrometry as an authentication method for mammalian cell lines of common genetic origin.
Main Methods:
- Intact cell MALDI-ToF mass spectrometry was applied to three parental neuroblastoma cell lines (IMR-5, IMR-32, UKF-NB-3) and eleven drug-adapted sublines.
- Principal component analysis (PCA) was used to analyze the mass spectrometry data and identify differences between cell lines.
- Whole-cell mass spectrometry fingerprints were generated for each cell line.
Main Results:
- PCA of MALDI-ToF mass spectrometry data revealed distinct patterns differentiating most, but not all, cell lines.
- The method successfully separated the parental cell line IMR-32 from its clonal subline IMR-5.
- Drug-adapted sublines, including cisplatin/oxaliplatin-resistant and vincristine/vinblastine-adapted lines, exhibited unique mass spectrometry patterns.
Conclusions:
- Intact whole-cell MALDI-ToF mass spectrometry demonstrates potential for distinguishing between cell lines with a common genetic background.
- This technique could be developed into a valuable tool for cell line authentication, complementing existing methods.
- The findings suggest MALDI-ToF mass spectrometry can identify subtle molecular differences arising from drug adaptation or clonal variation.
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